Renovación, proyecto arquitectónico y apropiación

Proyecto Terminal (Licenciatura en Sociología) -- Universidad Autónoma Metropolitana, Unidad Azcapotzalco, División de Ciencias Sociales y Humanidades, Departamento de Sociología, 1989. UAMADCSHDS . 1 archivo PDF (123 páginas)El Programa Emergente de Renovación Habitacional Popular, de la ciudad de México, por el cual muchas de las vecindades localizadas en el centro fueron transformadas en condominios en donde los inquilinos pasaron a ser propietarios de una vivienda. Da pie a este trabajo donde se estudiará el cambio cultural ocasionado por: la transformación física de la vivienda, cambio jurídico, gastos e implicaciones de la nueva vivienda; nuevas elaciones sociales originadas por la guía de autoadministración de propiedad en condominio de carácter vecinal, el uso de espacios privados, y comunes e influencia de estos en las relaciones sociales. El estudio se realiza en la colonia Morelos

Papel del Estroma Medular en la Mejora de la Función Hematopoyética: Desde la Irradiación hasta las Vesículas Extracelulares

[EN] Background Bone marrow (BM) microenvironment regulates growth and differentiation of hematopoietic stem cells (HSC) and is composed of several cell types, including osteoblasts (that have a critical role in the regulation of hematopoiesis and in the maintenance of the clonogenic potential of HSC) and adipocytes (which exert an inhibitory effect on hematopoiesis), both derived from mesenchymal stromal cells (MSC). Allogeneic hematopoietic cell transplantation (allo-SCT) remains as the only curative therapeutic approach for a variety of hematopoietic diseases. An adequate hematopoietic function after an allo-SCT is not only dependent on the number of hematopoietic stem cells infused, that is a key factor for engraftment. In the last few years, the focus has also been oriented to the BM microenvironment, especially to MSC. In the current work, we have hypothesized that inducing some modifications in the BM microenvironment (as the ones induced by low-dose irradiation, that is increasingly used in the conditioning regimens used in elderly patients) or the release of MSC-derived extracellular vesicles (MSC-EV) (one of the most important mechanism by which MSC exert their therapeutic effects) could increase the hematopoietic function, with its potential application to improve the outcome of allo-SCT, especially in those cases with poor graft function. Objectives Thus, the main objective of our work was to evaluate multiparametrically the effects of a reduced dose (2.5Gy) of irradiation on MSC and the effects of the incorporation of MSC-EV into HSC and their potential role in the improvement of the hematopoietic function. The specific aims were: 1) To evaluate the effects of low-dose irradiation on MSC and their impact on hematopoietic function, 2) To study the effects on MSC-EV incorporation into CD34+ cells and their impact on hematopoietic function, 3) To study the effects of previous-irradiation of MSC on their VE (IRR-MSC-EV) capacity on hematopoietic function improvement, and 4) To evaluate the effects of different doses of MSC-EV incorporation into HSC and their capacity on hematopoietic function improvement. Methods In the current work, a total of 70 bone marrow samples from healthy donors, 45 leukapheresis samples and 33 cord blood units were used, after proper informed consent was obtained and with the approval of the local Ethics Committee. To evaluate the effects of low-dose irradiation on MSC and their impact on hematopoietic function, MSC at third passage were irradiated with 2.5 Gy or not (the latter were used as controls). Cells were characterized following International Society for Cellular Therapy criteria, including in vitro differentiation assays. Adipogenic differentiation was assessed by Oil-Red staining and reverse transcriptase (RT)-PCR of CEBPA and PPARG, osteogenic differentiation was evaluated by alkaline phosphatase staining and RT-PCR of RUNX2 and ALP and mineralization was analyzed RT-PCR of SPP1 and quantified by Alizarin Red staining. Apoptosis was evaluated by flow cytometry with annexin V/7-AAD staining. Gene expression profile was studied by Chip Human Gene ST Arrays and the most relevant genes involved in hematopoiesis maintenance (SDF-1, ANGPT-1, COL1A1, THPO, CXCR4, ITGA-4, CD44 and NGF) were analyzed by RT-PCR, SDF-1 expression was confirmed by ELISA. Finally, long-term bone marrow cultures were performed to test the hematopoietic-supporting ability. Clonal growth of progenitor cell population was assayed weekly culturing CD34+ cells in methylcellulose Media. Differentiation status of stroma was evaluated during culture. To study the effects on MSC-EV incorporation into CD34+ cells and their impact on hematopoietic function, MSC-EV were characterized by flow cytometry, Western blot, electron microscopy (TEM), and nano-particle tracking analysis (NTA). Micro-RNA content of MSC-EV and IRR-MSC-EV was analyzed by TaqMan Arrays. 1x105 CD34+ cells were co-cultured with EV isolated from 3x106 MSC and EV incorporation into CD34+ cells was confirmed by flow cytometry and confocal microscopy after staining EV with Vybrant Dil cell labeling solution. Then Gene expression profile was studied by Chip Human Gene ST Arrays. Apoptosis and cell cycle were evaluated by flow cytometry and Caspase 3/7 and Caspase 9 activity was measured by luminescence. RT-PCR were performed in modified CD34+ cells in order to analyze expression of some genes (SDF-1, COL1A1, CD44, CXCR4, ITGA-4 y cKIT) and micro-RNAs (150, 155, 181a, 17, 363, 494 and Let7g). Protein expression of CD44, CXCR4, ITGA-4 and cKIT was evaluated by flow cytometry and CXCR4 and cKIT expression was confirmed by Western blot (Wes Simple). Phosphorylation of STAT5 was also analyzed by WES Simple. Finally, clonal growth of CD34+ cells in the different experimental conditions (after MSC-EV incorporation or not) was assessed by clonogenic assays and their capacity of engraftment was analyzed 4 weeks after CD34+ cell transplantation in non-obese diabetic/severe combined immunodeficient mice by flow cytometry in bone marrow and spleen. Similar experiments were done to evaluate either the effect of different doses of MSC-EV (isolated from 3x106MSC or 9x106MSC), or the effect of EV released by low-dose irradiated MSC on CD34+ cells. Results After low-dose irradiation of MSC, the immunophenotypic characterization and viability of irradiated MSC was comparable to that of control cells. Gene expression profiling showed a significant differential expression in 50 genes. Of them, 5 genes were overexpressed and 45 were down-regulated in irradiated compared with non-irradiated MSC. The most downregulated gene was pyruvate dehydrogenase kinase 1 (PDK1), which is involved in the regulation of adipogenesis. By RT-PCR, we observed that SDF-1 and ANGPT were overexpressed, whereas COL1A1 was down-regulated in irradiated cells (p=0.015, p=0.007, and p=0.031, respectively). The over-expression of SDF-1 in irradiated cells was confirmed by ELISA. Analyzing their differentiation capacity, we observed that, differentiation of irradiated MSC was skewed toward osteogenesis, whereas adipogenesis was impaired. Higher expression of SPP1 (p=0.039), involved in mineralization, and lower expression of genes involved in adipogenesis, CEBPA and PPARG (p=0.003 and p=0.019), was observed in irradiated cells. Moreover, an increase in the mineralization capacity quantified by Alizarin Red staining and a decrease in adipocyte counts were observed in irradiated cells at days 7, 14, and 21 after culture in specific differentiation media (p=0.018 p=0.046, and p=0.018, respectively). Finally, colony-forming unit granulocyte macrophage (CFU-GM) numbers in long-term bone marrow cultures were higher in the irradiated cells during the five weeks of the culture, with significant differences after 4 and 5 weeks (p=0.046 and p=0.007). In summary, the irradiation of MSC with 2.5 Gy improved their hematopoietic-supporting ability by increasing osteogenic differentiation and decreasing adipogenesis. Regarding the effects on MSC-EV incorporation into CD34+ cells and their impact on hematopoietic function, the isolation and characterization of the EV showed that EV size evaluated by NTA was homogeneous among samples with a mean of 131.93 nm (124.4–143.6 nm) and a mean particle concentration of 9.09E+10 particles/milliliter (5.16E+10–1.21E+11) in preparations of EV isolated from 3x106MSC. In addition, by TEM we observed the characteristic rounded morphology of EV. By flow cytometry, we confirmed that EV were smaller than 1 μm, negative for hematopoietic markers and positive for MSC and exosome markers (CD81 and CD63). CD63 expression was also confirmed by Western blot. MSC-EV incorporation was visualized by confocal microscopy and quantified by flow cytometry. Upon incorporation into CD34+ cells, MSC-EV induced a down-regulation of proapoptotic genes, an overexpression of genes involved in colony formation, and an activation of the JAK-STAT pathway. A significant decrease in apoptosis and Caspases 3/7 and Caspase 9 activation was observed in CD34+ cells after the incorporation of MSC-EV. Increased CD44 and CXCR4 expression and decreased cKIT expression upon the incorporation of MSC-EV were confirmed by FC. Increased levels of phospho-STAT5 were detected by WES Simple in CD34+ cells with MSC-EV. In addition, these cells displayed a higher colony-forming unit granulocyte/macrophage clonogenic potential and the in vivo bone marrow lodging ability of human CD34+ cells with MSC-EV was significantly increased in the injected femurs. In summary, the incorporation of MSC-EV induces genomic and functional changes in CD34+ cells, increasing their clonogenic capacity and their bone marrow lodging ability. When comparing the effects of EV from pre-irradiated MSC (IRR-MSC-EV) we observed that were similar in size, morphology, concentration and immunophenotype to non irradiated MSC-EV and they had similar capacity of incorporation into CD34+ cells. Regarding micro-RNA content, we found 19 micro-RNAs significantly downregulated in IRR-MSC-EV compared to MSC-EV. Some of them were analyzed in CD34+ cells that had incorporated either MSC-EV or IRR-MSC-EV but we did not found differences in its expression in these cells. However, upon the incorporation of IRR-MSC-EV, 1330 genes were modified in CD34+ cells inducing a downregulation of proapoptotic genes, an overexpression of genes involved in colony formation, and an activation of the JAK-STAT pathway as MSC-EV did. Moreover, we detected an over-expression of many HLA molecules resulting in the overexpression of antigen processing and presentation, graft versus host disease or allograft rejection pathways. Also proteasome, ribosome biogenesis and other pathway were altered. Despite these differences in gene expression, we did not find differences in viability, cell cycle, expression of molecules involved in hematopoiesis or colony formation capacity between CD34+ cells that had incorporated IRR-MSC-EV or MSC-EV. In addition, IRR-MSC-EV and MSC-EV significantly increase BM lodging ability of CD34+ cells without significant differences among both experimental groups. Finally we evaluated the potential impact of the MSC-EV dose in the observed effects in CD34+ cells. As expected, particle concentration assessed by NTA of EV preparations isolated from 9x106 MSC (3.02E+12 particles/ml) was higher than that of EV isolated from 3x106MSC (2,88E+11 particles/ml). Besides, we observed by flow cytometry a significant increase of CD34+ cells that had incorporated EV when they were co-cultured with the higher dose of EV. However, despite the fact that some effects (increase of viability, CD44 expression and clonogenic capacity) were more pronounced after the incorporation of higher dose of EV in CD34+ cells, we did not found significant differences between different doses, neither in their bone marrow lodging ability. So we can conclude that the lower dose (VE isolated from 3x106 MSC) could be optimal for inducing changes in CD34+ cells in the experiments performed and that higher doses do not contribute to improve these beneficial effects. Conclusions 1) Low-dose γ-irradiation of MSC causes an alteration in their gene expression profile increasing the expression levels of SDF-1 and ANGPT, favoring osteogenic differentiation and decreasing adipogenesis. These modifications lead to an improvement of MSC hematopoietic-supporting ability. 2) Human MSC-EV are able to incorporate into human CD34+ cells, modifying their gene expression and increasing their viability, clonogenic capacity in vitro, and their 4-week BM lodging ability in vivo. 3) IRR-MSC-EV induce similar genomic and functional changes as MSC-EV when incorporate into CD34+ cells without significant differences in the viability, cell cycle, colony forming capacity and BM lodging ability of CD34+ cells. These results confirms that the beneficial effect of low-irradiation on MSC hematopoietic-supporting ability is not related to their exchange of molecules through EV. 4) EV isolated from 9x106 MSC do not improve the beneficial effects in the hematopoietic function induced by EV isolated from 3x106 when they incorporate into CD34+ cells

Uso de canciones como recurso didáctico para mejorar la capacidad de comprensión y expresión oral del idioma inglés en los estudiantes del VI ciclo de educación secundaria de la I.E. “Santiago Antúnez de Mayolo”, Santa - 2017

El presente informe de investigación denominado USO DE CANCIONES COMO RECURSO DIDÁCTICO PARA MEJORAR LA CAPACIDAD DE COMPRENSIÓN Y EXPRESIÓN ORAL DEL IDIOMA INGLÉS EN LOS ESTUDIANTES DEL VI CICLO DE EDUCACIÓN SECUNDARIA DE LA I.E. “SANTIAGO ANTÚNEZ DE MAYOLO”, SANTA - 2017, fue desarrollado con el objetivo de demostrar que las canciones en inglés mejoran la capacidad de comprensión y expresión oral de este idioma en los estudiantes del VI ciclo de educación secundaria. La investigación fue de tipo experimental con un diseño de investigación cuasi- experimental. La población del estudio estuvo compuesta por 66 estudiantes, asimismo se eligió la sección del primer año “A” como grupo experimental conformado por 23 estudiantes y la sección “B” como grupo de control conformado por 21 estudiantes. A ambos grupos se les aplicó un pre-test y post-test, los cuales permitieron la recolección de datos para la comprensión oral y el uso de la rúbrica como instrumento de evaluación para la expresión oral. La hipótesis central de la investigación fue que las canciones como recurso didáctico mejora la competencia comunicativa. Los resultados demostraron que el uso de las canciones en inglés si mejoró la capacidad de comprensión y expresión oral pues generó un clima agradable en el aula, redujo el estrés, ofreció un ambiente agradable para desenvolverse en la lengua meta y motivar a los estudiantes a ser partícipes de su propio aprendizaje.Tesi

What if students propose their own examinations? An introductory experiment

[EN] This paper presents a proposal experimented in the University of Burgos, where students of different degrees, at a Bachelor or Master level, had to prepare solved questions or exercises, in order to introduce some of them into the exams. There were considered three types of experiment: test, short questions and exercises. Students submitted they own proposals into forum, where the teacher corrected them and randomly selected some of them, which configured the half of the official exam. The obtained results were very positive: students had a very good perception of the experiment, learnt more and passed the subject easily. Also, the average grading were higher that the corresponding with the classical system, existing a slight correlation between this increase and their point of view about the chance to learn. The satisfaction and the marks of the students are higher, while having a better knowledge of the topics included. Also, students have the feeling that the teachers are more considerate and thoughtful with them, so this type of experiments should be continued.Rojo, M.; Preciado, M.; Gonzalo-Orden, H.; Moreno, I.; Casado, S.; Cárdenas, D. (2017). What if students propose their own examinations? An introductory experiment. En Proceedings of the 3rd International Conference on Higher Education Advances. Editorial Universitat Politècnica de València. 566-573. https://doi.org/10.4995/HEAD17.2017.5290OCS56657

Predictores de eventos clínicos en una cohorte de pacientes con enfermedad de Fabry en tratamiento sustitutivo enzimático

Enfermedad de Fabry; Enfermedad renal crónica; Tratamiento enzimático sustitutivoFabry disease; Chronic kidney disease; Enzyme replacement therapyMalaltia de Fabry; Malaltia renal crònica; Tractament enzimàtic substitutiuFabry disease may be treated by enzyme replacement therapy (ERT), but the impact of chronic kidney disease (CKD) on the response to therapy remains unclear. The aim of the present study was to analyse the incidence and predictors of clinical events in patients on ERT. Study design: Multicentre retrospective observational analysis of patients diagnosed and treated with ERT for Fabry disease. The primary outcome was the first renal, neurological or cardiological events or death during a follow-up of 60 months (24-120). Results: In 69 patients (42 males, 27 females, mean age 44.6±13.7 years), at the end of follow-up, eGFR and the left ventricular septum thickness remained stable and the urinary albumin: creatinine ratio tended to decrease, but this decrease only approached significance in patients on agalsidase-beta (242-128mg/g (p=0.05). At the end of follow-up, 21 (30%) patients had suffered an incident clinical event: 6 renal, 2 neurological and 13 cardiological (including 3 deaths). Events were more frequent in patients with baseline eGFR≤60ml/min/1.73m2 (log Rank 12.423, p=0.001), and this remained significant even after excluding incident renal events (log Rank 4.086, p=0.043) and in males and in females. Lower baseline eGFR was associated with a 3- to 7-fold increase the risk of clinical events in different Cox models. Conclusions: GFR at the initiation of ERT is the main predictor of clinical events, both in males and in females, suggesting that start of ERT prior to the development of CKD is associated with better outcomes.El objetivo de este estudio es realizar un mapa del tratamiento actual de la enfermedad de Fabry en España, analizando el efecto de diferentes factores en el desarrollo de eventos clínicos a largo plazo. Diseño del estudio: Análisis observacional retrospectivo multicéntrico. Criterios de inclusión: pacientes diagnosticados y tratados de enfermedad de Fabry. Se recogieron datos generales en relación con el diagnóstico, síntomas y tipo mutación, tipo de tratamiento recibido, evolución renal y cardiológica. Durante un tiempo de seguimiento de 60 meses (24-120), se recogió el primer evento clínico tras el inicio de tratamiento sustitutivo enzimático definido como mortalidad, evento renal, cardiológico o neurológico. Resultados: Se incluyeron 69 pacientes (42 H, 27 M) con una edad media de 44,6 ± 13,7 años. A los cinco años de tratamiento, el FGe y la hipertrofia ventricular izquierda se mantuvieron estables, y la albuminuria tiende a disminuir, siendo este descenso más significativo en el grupo de pacientes tratados con beta-galactosidasa (de 242 a 128mg/g (p = 0,05). Veintiún pacientes sufrieron un evento clínico (30%): seis renales, dos neurológicos y 13 cardiológicos (incluidas tres muertes). Los pacientes con ERC (FGe < 60) antes del inicio de tratamiento tuvieron más eventos (log-rank 12.423, p = 0,001), manteniéndose la predicción si excluíamos los eventos renales (log-rank 4.086 (p = 0,043) en hombres y mujeres. La peor función renal al inicio del tratamiento aumentó entre tres y siete veces el riesgo de eventos clínicos en diferentes modelos de Cox ajustados. Conclusiones: La función renal al inicio de tratamiento sustitutivo enzimático es la principal predictora de desarrollo de eventos clínicos a largo plazo, tanto en hombres como mujeres. El inicio de tratamiento sustitutivo enzimático precoz antes del desarrollo de ERC mejoraría el pronóstico.MG, JT, AO, RT are supported by ISCIII RETIC REDINREN, RD016/009 and FEDER fund

Co-administration of human MSC overexpressing HIF-1α increases human CD34+ cell engraftment in vivo

Background: Poor graft function or graft failure after allogeneic stem cell transplantation is an unmet medical need, in which mesenchymal stromal cells (MSC) constitute an attractive potential therapeutic approach. Hypoxia-inducible factor-1α (HIF-1α) overexpression in MSC (HIF-MSC) potentiates the angiogenic and immunomodulatory properties of these cells, so we hypothesized that co-transplantation of MSC-HIF with CD34+ human cord blood cells would also enhance hematopoietic stem cell engraftment and function both in vitro and in vivo. Methods: Human MSC were obtained from dental pulp. Lentiviral overexpression of HIF-1α was performed transducing cells with pWPI-green fluorescent protein (GFP) (MSC WT) or pWPI-HIF-1α-GFP (HIF-MSC) expression vectors. Human cord blood CD34+ cells were co-cultured with MSC WT or HIF-MSC (4:1) for 72 h. Then, viability (Annexin V and 7-AAD), cell cycle, ROS expression and immunophenotyping of key molecules involved in engraftment (CXCR4, CD34, ITGA4, c-KIT) were evaluated by flow cytometry in CD34+ cells. In addition, CD34+ cells clonal expansion was analyzed by clonogenic assays. Finally, in vivo engraftment was measured by flow cytometry 4-weeks after CD34+ cell transplantation with or without intrabone MSC WT or HIF-MSC in NOD/SCID mice. Results: We did not observe significant differences in viability, cell cycle and ROS expression between CD34+ cells co-cultured with MSC WT or HIF-MSC. Nevertheless, a significant increase in CD34, CXCR4 and ITGA4 expression (p = 0.009; p = 0.001; p = 0.013, respectively) was observed in CD34+ cells co-cultured with HIF-MSC compared to MSC WT. In addition, CD34+ cells cultured with HIF-MSC displayed a higher CFU-GM clonogenic potential than those cultured with MSC WT (p = 0.048). We also observed a significant increase in CD34+ cells engraftment ability when they were co-transplanted with HIF-MSC compared to CD34+ co-transplanted with MSC WT (p = 0.016) or alone (p = 0.015) in both the injected and contralateral femurs (p = 0.024, p = 0.008 respectively). Conclusions: Co-transplantation of human CD34+ cells with HIF-MSC enhances cell engraftment in vivo. This is probably due to the ability of HIF-MSC to increase clonogenic capacity of hematopoietic cells and to induce the expression of adhesion molecules involved in graft survival in the hematopoietic niche

Nanopartículas de quitosán mejoran el rendimiento, actividad enzimática y compuestos bioactivos en frutos de tomate

Las nanopartículas de quitosán (NPsCS) son utilizadas como bioestimulantes naturales en la agricultura sustentable, ya que incrementan la productividad de los cultivos e inducen la síntesis de antioxidantes enzimáticos y no enzimáticos, protegiendo a la planta del estrés. El presente estudio se desarrolló con el objetivo de determinar el efecto de la aplicación foliar de las NPsCS sobre el rendimiento, actividad enzimática y contenido de compuestos bioactivos en frutos de tomate. El ensayo se estableció en un diseño completamente al azar con seis dosis crecientes de NPsCS: 0, 0.05, 0.1, 0.2, 0.4 y 0.8 mg mL-1. La aspersión foliar de 0.2 mg mL-1 aumentó el rendimiento, tamaño y firmeza de los frutos; en cambio dosis alta incrementan los compuestos bioactivos y la actividad enzimática. El uso de NPsCS aplicadas de forma foliar presentan un gran potencial para utilizarse como bioestimulantes para mejorar el rendimiento y obtener alimentos funcionales

Experiencia piloto de “micro flipped teaching” (clase invertida) en la asignatura “Células madre de la Médula Ósea: características biológicas y su posible papel en el desarrollo de neoplasias”

Memoria ID2019/067. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2019-2020

Mesenchymal stromal cells combined with elastin-like recombinamers increase angiogenesis in vivo after hindlimb ischemia

Producción CientíficaHindlimb ischemia is an unmet medical need, especially for those patients unable to undergo vascular surgery. Cellular therapy, mainly through mesenchymal stromal cell (MSC) administration, may be a potentially attractive approach in this setting. In the current work, we aimed to assess the potential of the combination of MSCs with a proangiogenic elastin-like recombinamer (ELR)–based hydrogel in a hindlimb ischemia murine model. Human bone marrow MSCs were isolated from four healthy donors, while ELR biomaterials were genetically engineered. Hindlimb ischemia was induced through ligation of the right femoral artery, and mice were intramuscularly injected with ELR biomaterial, 0.5 × 106 MSCs or the combination, and also compared to untreated animals. Tissue perfusion was monitored using laser Doppler perfusion imaging. Histological analysis of hindlimbs was performed after hematoxylin and eosin staining. Immunofluorescence with anti–human mitochondria antibody was used for human MSC detection, and the biomaterial was detected by elastin staining. To analyze the capillary density, immunostaining with an anti–CD31 antibody was performed. Our results show that the injection of MSCs significantly improves tissue reperfusion from day 7 (p = 0.0044) to day 21 (p = 0.0216), similar to the infusion of MSC + ELR (p = 0.0038, p = 0.0014), without significant differences between both groups. After histological evaluation, ELR hydrogels induced minimal inflammation in the injection sites, showing biocompatibility. MSCs persisted with the biomaterial after 21 days, both in vitro and in vivo. Finally, we observed a higher blood vessel density when mice were treated with MSCs compared to control (p<0.0001), but this effect was maximized and significantly different to the remaining experimental conditions when mice were treated with the combination of MSCs and the ELR biomaterial (p < 0.0001). In summary, the combination of an ELR-based hydrogel with MSCs may improve the angiogenic effects of both strategies on revascularization of ischemic tissues.Spanish Government (RTI2018-096320-B-C22, FPU16-04015, PID2019-110709RB-I00, PID2020-118669RA-I00)Interreg V España Portugal POCTEP (0624_2IQBIONEURO_6_E), Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y LeónConsejería de Educación de Castilla y León (CAS079P17)Instituto de Salud Carlos III (ISCIII) (PI19/01630)Programs of ISCIII- European Regional Development Fund (RD16/0011/0015, RD21/ 0017/0006